The Effect of Subjecting Peas to Air Enriched with Carbon Dioxide: II. RESPIRATION AND THE METABOLISM OF THE MAJOR ACIDS

Whole peas at about 75 per cent of their maximum fresh weightwere subjected to 5–30 per cent CO₂ in air for periodsof from 1–6 d, then returned to air for a further 1 d.Samples were withdrawn at intervals and organic acids, ^TCO₂and ethanol estimated as well as the rate of respiration. Slicesof cotyledons suspended in water were also subjected to highconcentrations of CO₂ in air for 3 h. The rate of respiration was inhibited progressively by increasein CO₂ content of the tissue. The high internal CO₂ contentof the intact pea causes an inhibition of its rate of respirationby about 25 per cent. Alcohol production commenced at between10 and 15 per cent CO₂ in the ambient gas and slowly increasedin rate up to 37 per cent. The CO₂-air mixtures reduced the content of malate, pyruvateand -oxoglutarate, increased that of succinate and left citrateunaffected. On return to air malate rose rapidly and succinatefell slowly to their original concentrations. During the sameperiod the concentration of PEP fell sharply and after about1 h rose again, whereas oxalacetate showed a reverse response.It is argued that the rapid re-synthesis of malate was by carboxylationof PEP to oxalacetate and that this reaction was stimulatedby a change in pH rather than by the direct effect of the changein concentration of CO₂. In one experiment ¹⁴CO₂ was supplied for 2 h before return toair and the movement of ¹⁴C followed for 6 h. The results supportthe method of re-synthesis of malate proposed.     

STUDIES ON PERMEABILITY OF MEMBRANES : I. INTRODUCTION AND THE DIFFUSION OF IONS ACROSS THE DRIED COLLODION MEMBRANE.

The theoretical aspects of the problem of sieve-like membranes are developed. The method of preparing the dried collodion membrane is described, and the method of defining the property of a particular membrane is given. It consists of the measurement of the Co P, that is the P.D. between an 0.1 and an 0.01 M KCl solution separated by the membrane. Co P is in the best dried membranes 50 to 53 millvolts, the theoretically possible maximum value being 55 millivolts. Diffusion experiments have been carried out with several arrangements, one of which is, for example, the diffusion of 0.1 M KNO₃ against 0.1 M NaCl across the membrane. The amount of K⁺ diffusing after a certain period was in membranes with a sufficiently high Co P (about 50 millivolts or more) on the average ten times as much as the amount of diffused Cl^-. In membranes with a lower Co P the ratio was much smaller, down almost to the proportion of 1:1 which holds for the mobility of these two ions in a free aqueous solution. When higher concentrations were used, e.g. 0.5 M solution, the difference of the rate of diffusion for K⁺ and Cl^- was much smaller even in the best membranes, corresponding to the fact that the P.D. of two KCl solutions whose concentrations are 10:1 is much smaller in higher ranges of concentration than in lower ones. These observations are confirmed by experiments arranged in other ways. It has been shown that, in general, the diffusion of an anion is much slower than the one of a cation across the dried collodion membrane. The ratio of the two diffusion coefficients would be expected to be calculable in connection with the potential difference of such a membrane when interposed between these solutions. The next problem is to show in how far this can be confirmed quantitatively.     

STUDIES ON THE BIOSYNTHESIS OF COLLAGEN : I. THE GROWTH OF FOWL OSTEOBLASTS AND THE FORMATION OF COLLAGEN IN TISSUE CULTURE

1. A tissue culture method was devised in which suspensions of osteoblasts, obtained directly from frontal bones of fowl embryos, were grown in a fluid, fibrin-free medium. 2. Maximum growth of the tissue, as measured by dry weight, with the formation of collagen protein, based on the estimation of hydroxyproline, was obtained in periods of up to 6 days. 3. Appreciable amounts of protein-bound hydroxyproline were formed during the first 24 hour growth period, but electron microscopy of portions of the same cultures failed to demonstrate the presence of any typical collagen fibrils. 4. The subsequent formation of many characteristic collagen fibrils was not associated with a significant rise in the mean hydroxyproline content of the tissue. 5. The cytoplasmic granules of the osteoblasts stained intensely with the P.A.S. technique when the collagen fibrils were being formed. 6. It is suggested that collagen-forming cells synthesise and secrete a hydroxyproline-rich precursor of protein or large peptide nature, which subsequently becomes directly transformed into typical collagen fibrils.     

Dormancy in Rice Seed: III. THE INFLUENCE OF TEMPERATURE, MOISTURE, AND GASEOUS ENVIRONMENT

The influence of storage conditions—temperature, moisturecontent, oxygen, nitrogen, and carbon dioxide—on dormancyin rice seed has been investigated. The effects of temperatureand oxygen when the seeds have been set to germinate have alsobeen studied. Storage in oxygen accelerates the breaking of dormancy; at lowertemperatures, the effect of oxygen is more noticeable. Carbondioxide and nitrogen have little or no effect except in so faras they exclude oxygen. It is shown that there is a negativelinear relationship between storage temperature and log. meandormancy period of intact seeds over the range 27°C. to47°C. In the variety tested, the Q₁₀ for the rate of breakingdormancy is 3.38. Variation of moisture content over the range12.0–14.5 per cent, has little effect at 27°C. andno detectable effect at higher temperatures. When dormant seeds are soaked in water, they attain a moisturecontent close to 30 per cent. Pre-soaking seeds to achieve moisturecontents in this region can stimulate the breaking of dormancywhen compared with dry storage. At laboratory temperature (meanabout 27°C.) the stimulation caused by pre-soaking intactseeds is usually small and sometimes non-existent, but at alow temperature (3°C.) the stimulation is increased; butdry storage at 3°C. markedly delays the breaking of dormancy.When seeds are dehusked, a large proportion lose their dormancy.Much of the residual dormancy of dehusked seeds can be brokenby soaking at laboratory temperature. But in the sample of dehuskedseeds used, low temperature did not increase the effect of thepre-soaking, but slightly decreased the stimulation caused bythe soaking treatment in this case. The effect of temperature on wet seed has also been investigatedwhen the seed has been set to germinate. Maximum germinationof a partially dormant population of seed is achieved at anoptimum temperature which is near or somewhat below 27°C.In the sample tested, no germination occurred at 42°C.,although in populations which have completely broken dormancyfull germination will occur at this temperature. Non-dormantpopulations of rice seed can germinate at very low oxygen tensionsor probably even in the absence of oxygen, but germination ofa partially dormant sample is reduced under these circumstances.When oxygen tension is very low there is less germination at27°C. than at 37°C. In some varieties, at least, there is evidence that the seedsgo through a stage when they will germinate in daylight butnot in the dark. A simple practical method for rapidly breakingthe dormancy of intact seeds is described. The significanceof these results is discussed in relation to the published workon seeds of other species.     

BIOGENESIS OF ETHYLENE IN APPLE TISSUE: I. FORMATION OF ETHYLENE FROM GLUCOSE, ACETATE, PYRUVATE, AND ACETALDEHYDE IN APPLE TISSUE

1. With the aim of elucidating the path of carbon in the formationof ethylene in plants, studies were made on the incorporationof ¹⁴C into ethylene evolved from apple slices, using several¹⁴C- labeled compounds as substrates. The effects of inhibitorswere also investigated. 2. The formation of ethylene-¹⁴C from glucose-¹⁴C was inhibitedby fluoride, but unaffected by arsenite, thus suggesting thatglucose is converted to ethylene via pyruvate. 3. Acetate is converted to ethylene after cleavage of C-l andC-2. Only a small portion of the latter (C-2) enters the moleculeof ethylene, the former (C-l) is detected in carbon dioxide.On the other hand, 2, and 3-carbons of pyruvate are converted,without splitting, to ethylene. 4. On removal of air, the incorporation of ¹⁴C into ethylenefrom acetate-2-¹⁴C was depressed, while that from pyruvate-¹⁴Cwas unaffected. 5. Acetaldehyde-l,2-¹⁴C is converted to ethylene without conversioninto ethanol. 6. These results are interpreted to suggest the occurrence ofthe pathway in which pyruvate and acetaldehyde may serve asprecursors of ethylene. ¹ A part of this paper was read at the regular Meeting of KansaiBranch of the Agricultural Chemical Society of Japan in Kyoto,October, 1964, and at the Annual Meeting of the Japanese Societyof Plant Physiologists in Tokyo, April, 1965 and presented ina preliminary form elsewhere (10).     

The Initiation and Development of Lateral Meristems in the Pea Root: I. THE EFFECT OF YOUNG AND OF MATURE TISSUE

The effect of.distinct regions of the root on the initiationof lateral root primordia and the emergence of lateral rootshas been studied, using segments of roots from sterile 2-daygerminated pea seedlings. It is shown that the removal of the basal region causes a decreasein the number of primordia formed in the remainder of the root.On the other hand, the removal of the apical region causes alarger number of primordia to be formed in the remaining tissuethan in the corresponding tissue of roots where the apical regionis retained. It is suggested that a factor or a complex of factorsinvolved in primordium initiation is translocated from the oldertissue towards the potential site of primordium initiation inthe young tissue which has just completed extension growth. The removal of the apical region of the root is also shown tostimulate lateral root emergence. It is suggested that a factoror complex of factors involved in the development of the primordiasubsequent to initiation moves within the root in a similarmanner to the factor or factors involved in initiation.     

Devlopmental Physiology of Sugar-Beet: II. EFFECTS OF TEMPERATURE AND NITROGEN SUPPLY ON THE GROWTH, SOLUBLE CARBOHYDRATE CONTENT AND NITROGEN CONTENT OF LEAVES AND ROOTS

Plants were grown at temperatures of 15 and 25 ?C with two ratesof nitrogen supply. The changes in dry weight, leaf area, cellnumber, mean cell volume, soluble carbohydrate, and total nitrogenconcentration of the cotyledons, the first and second pair oftrue leaves, and the storage root were measured. Changes incell number and cell volume of the first pair of true leavesand storage root of plants were also measured at 11, 18, 25,and 32 ?C. Leaf growth before unfolding was chiefly by increase in cellnumber and after unfolding by increase in mean cell volume,while the growth of the storage root was almost entirely byincrease in cell number. The rates of cell division and cellexpansion were fastest at 25 ?C, but the initially high ratesof cell division in the terminal bud and in individual leavesdecreased rapidly and greater rates were maintained at the sub-optimaltemperatures, i.e. 15 and 18 ?C. After an initial period ofslow growth, the first-formed leaves grew faster and becamelarger at 15 than at 25 ?C. Leaves were produced, unfolded,grew faster, and became larger with increase in the externalconcentration of nitrogen, because cells divided and expandedfaster, so that nitrogen increased the number and size of cells. Sugar concentration was greater at 15 than at 25 ?C in leavesbut not in the storage root. Sugar concentration in the petiolesof the first and second pair of true leaves increased to 1.2and 2.0 per cent fresh weight respectively. Decreased nitrogensupply temporarily increased the sugar concentration of cotyledonpetioles and the seedling hypocotyl, but later decreased itin the leaves and storage root. Nitrogen concentration was greaterin the leaves and storage root at 15 than at 25 ?C with thelarger nitrogen supply. Nitrogen concentrations were similarin young leaves of all treatments but as the size of leavesincreased nitrogen concentrations decreased most rapidly at25 ?C with the smaller nitrogen supply. It is suggested that when increased leaf production and storage-rootgrowth occurs at temperatures below the growth optimum (25 ?C),they may be due to an effect of increased carbohydrate supplyon cell division and sugar storage.     

Translocation in Sugar-beet: I. ASSIMILATION OF 14CO2 AND DISTRIBUTION OF MATERIALS FROM LEAVES

¹⁴CO₂ was supplied to leaves, and movement of labelled carbonto other parts of the plant was assessed. Young growing leavesutilized assimilated carbon for their own growth and did notexport carbon to the rest of the plant, while fully expandedleaves exported much of their photosynthate, both to root andto young leaves. Translocation from a particular leaf was tothe two or three younger leaves on the same side of the plant,and to a sector of root below the source leaf. Specific distributionto growing leaves could be modified by partial defoliation.There was no movement of material to leaves which had emergedbefore the source leaf. Part of the carbon entering a leaf by assimilation (and, foryoung leaves, by translocation) was incorporated into insolublematerial, especially in young leaves. Some of the carbon enteringa developing root was permanently stored as sucrose, althoughmuch also entered insoluble material. Loss from the leaf ofcarbon fixed during a short period of photosynthesis was rapidat first but continued at a decreasing rate for several days.Some carbon fixed into the insoluble fraction was translocatedfrom the leaf later, during senescence. Sucrose was the mainmaterial translocated immediately after photosynthesis.     

The Transport and Metabolism of Urea in Chara australis: I. PASSIVE DIFFUSION, SPECIFIC TRANSPORT AND METABOLISM OF UREA, THIOUREA AND METHYLUREA

Most of the urea entering Chara australis cells is rapidly metabolizedto produce CO₂, which diffuses out of the cells into the surroundingmedium. A simple and convenient apparatus to measure both the¹⁴C-urea retained by cells and the ¹⁴CO₂ released into the mediumwas developed and used in a study of urea transport in Chara.The permeability coefficient for urea in the Chara plasmalemmawas estimated from the slope of an uptake versus concentrationfunction as 85 nm s^-1. Computer modelling of urea uptake andmetabolism suggests that this could be a 20% underestimate ofthe true value.The corresponding permeability coefficients forthiourea and N-methyl-urea were estimated in the same way as34 and 35 nm s^-1, respectively. These permeabilities are muchgreater than expected on the basis of either/water partitioncoefficients for the solutes and are consistent with the diffusionof urea and its similarly-sized analogues through aqueous poresin the plasmalemma.At external concentrations of urea less than20 mmol m^-3, the bulk of the uptake is effected by a specifictransport mechanism with an apparent K_m for urea of less than1.0 mmol m^-3. This transport system operates most rapidly withexternal pH in the range 6.5–7.5 and is influenced bythe nitrogen status of the cell.Evidence is produced here suggestingthat the specific transport of urea may be an active process. Key words: Chara, urea uptake, metabolism, diffusion, specific transport     

Water Relations of Tropical Epiphytes: I. RELATIONSHIPS BETWEEN STOMATAL RESISTANCE, RELATIVE WATER CONTENT AND THE COMPONENTS OF WATER POTENTIAL

The water relations of five species of tropical vascular epiphytesnative to Malaysia were studied. The species were ferns: Pyrrosiaadnascens (Forst.) Ching. and Pyrrosia angustata (Sw.) Ching.;orchids: Eria velutina Lindl., Dendrobium tortile Lindl. andDendrobium crumenatum Sw. Leaf resistance as a function of leafwater potential was measured for the two ferns. The criticalwater potential at which stomata closed was found to be highin each case; –0.75 MPa and –0.5 MPa respectively.The components of water potential were estimated with the pressurechamber as functions of relative water content. For each speciescell sap was found to be dilute, pressure potential low at fullturgor, and the change in relative water content between fullturgor and wilting point small. Small values of solute potentialat full turgor were also found for the ferns and E. velutinausing a vapour pressure osmometer. Values of the bulk modulusof elasticity of the leaf tissue for each species lay withinthe range of published data. The significance of these resultsfor the epiphytic way of life is discussed. Key words: Water potential, Epiphytes, Diffusive resistance, Orchid, Fern     

The Translocation of Antibiotics in Higher Plants: I. ISOLATION OF GRISEOFULVIN AND CHLORAMPHENICOL FROM PLANT TISSUE

Griseofulvin and chloramphenicol have been recovered from thetissues of broad-bean plants grown in solutions containing theseantibiotics. Partial separation of the compounds from other constituentsof solvent extracts of plant tissue was effected by counter-currentdistribution in a carbon tetra chloride methanol : water system.Pure griseofulvin and chloramphenicol were then obtained byrecovery of the solute in appropriate tubes followed by chromatographyon an activated alumina column; they were identified by mixedmelting point determinations and comparison of their infra-redspectra. A method for estimating griseofulvin in plant tissue extractsbased on counter current distribution and examination of thefractions spectrophotometrically has been developed, and usedto strike a balance between the gnseofulvin entering the plantand that present in the tissues after treatment. Estimates of chloramphenicol in the tissues by bioassay agreewell with the chemical determinations.     

SILICEOUS SCALE PRODUCTION IN CHRYSOPHYTE AND SYNUROPHYTE ALGAE. I. EFFECTS OF SILICA-LIMITED GROWTH ON CELL SILICA CONTENT,SCALE MORPHOLOGY,AND THE CONSTRUCTION OF THE SCALE LAYER OF SYNURA PETERSENII1

The cells of synurophyte flagellates (algal class Synurophyceae, formerly included in the Chrysophyceae) are enclosed within a regularly imbricate layer of ornamented siliceous scales. Scale morphology is of critical taxonomic importance within this group of algae, and the scales are valuable indicator microfossils in paleolimnological studies. The data presented here demonstrate that scale morphology and the integrity of the scale layer can exhibit extreme variability in culture as a function of the cellular quota of silica under silica-limited growth. Silica-limited, steady-state populations of the colonial flagellate Synura petersenii Korsh. were maintained over a range of specific growth rates (μ= 0.11–0.69 days^?1) and silica cell quotas (Q_si= 0.13–2.40 pmoles Si · cell¹). Scale morphology and the organization of the scale layer became increasingly aberrant as silica stress increased. Under severe stress, scale deposition was completely suppressed so that cells appeared scale-free. This depression of scale deposition was reversible; populations of silica-starved, scale-free cells rapidly regenerated new scale layers when placed in batch culture and spiked with dissolved silica. During recovery from silica stress, cell division was repressed for 24 h while mean cell silica quota increased 25-fold. The first new scales appeared within 2 h after the silica addition, and development of the new scale layer proceeded in an approximately synchronous manner, residting in normal scale layers on virtually all cells after 48 h of recovery in Sirich medium. Silica content of silica-replete Synura cells is comparable to freshwater diatoms of siynilar size, but Synura has much greater potential quota variability than diatoms and no apparent threshold silica requirement. Silica-limited growth kinetics and competition between diatoms and Synura for silica are discussed. The results suggest that morphological variability of siliceous scales in natural populations of synurophyte flagellates may result from silica stress and that the experimental approach developed here has great potential value as a means for circumscribing ecotypic variation in scale morphology. Results also demonstrate that scale production can be uncoupled from cell division, suggesting that cell cycle regulation of silica biomineralization in the Synurophyceae may be fundamentally different from that of diatoms (algal class Bacillariophyceae). This experimental system has application in the future study of the intracellular membrane systems and the regulatory processes involved in silica biomineralization.     

Carbon Dioxide Fixation in Soybean Roots and Nodules: I. CHARACTERIZATION AND COMPARISON WITH N(2) FIXATION AND COMPOSITION OF XYLEM EXUDATE DURING EARLY NODULE DEVELOPMENT

These studies demonstrate that soybean (Merr) roots and nodules possess an active system for fixing CO₂. The maximum rates of CO₂ fixation observed for roots and nodules of intact plants were 120 and 110 nanomoles CO₂ fixed per milligram dry weight per hour, respectively. Results of labeling studies suggest a primary role for phosphoenolpyruvate carboxylase in CO₂ assimilation in these tissues. After pulse-labeling with ¹⁴CO₂ for 2 minutes, 70% of the total radioactivity was lost within 18 minutes via respiration and/or translocation out of nodules. During the vegetative stages of growth of soybeans grown symbiotically, CO₂ fixation in nodules increased at the onset of N₂ fixation but declined to a lower level prior to the decrease in N₂ fixation. This decrease coincided with a decrease in the transport of amino acids, especially asparagine, and an increase in the export of ureides. These findings are consistent with a dual role for CO₂ fixation, providing substrates for energy-yielding metabolism and supplying carbon skeletons for NH₄⁺ assimilation and amino acid biosynthesis.     

Effects of CO2 Enrichment at Different Irradiances on Growth and Yield of Wheat: I. EFFECTS OF CULTIVAR AND OF DURATION OF CO2 ENRICHMENT

Wheat, Triticwn aestivum L., the winter cultivars Hobbit andCappelle-Desprez, and the spring cultivars Sicco and KJeiber,were grown in normal air or air enriched with CO₂ either outdoorsin a glass-roofed cage or in controlled environment rooms. Inneither the winter nor the spring wheat was growth increaseddue to enrichment with CO₂ before anthesis. Enrichment of thetwo winter wheat cultivars increased shoot dry weight significantlyat 15 d after anthesis but produced no significant increasein grain yield. With the spring cultivars there was a significantincrease in shoot dry weight by 18 d after anthesis and thegrain yield was also larger due to an increase in grain size.Shoot weight increased because the stems were larger, and therewas a diversion of assimilate from grain growth to late tillerproduction. Root tissue comprised less than 20% of the totaldry matter at anthesis (for all cultivars); effects of CO₂ enrichmenton root growth appeared to be less important than effects onshoot and ear growth. Growth and yield responses to CO₂ enrichmentwere observed (for the spring cultivars) at irradiances of both250 and 635 µE m^–2 s^–1, but the effects weregreater at the lower irradiance. Key words: CO₂ enrichment, Wheat, Cultivar     

The Geoelectric Effect in Plant Shoots: I. THE CHARACTERISTICS OF THE EFFECT

A comparative study has been made of the geoelectric effectin Helianthus hypocotyls and Zea coleoptiles using two electrodesystems. With the static-drop electrode system a potential differencedeveloped between the upper and lower surfaces of the shootsimmediately after they were turned into the horizontal position.The lower surface of the shoot became positively charged withrespect to the upper surface. In non-decapitated shoots thispotential difference continued to increase for at least 20 min,whereas in decapitated shoots no further increase occurred afterabout 10 min from the moment of reorientation. In contrast,with a flowing-solution electrode system no potential differencedeveloped in non-decapitated shoots until about 12 min afterthey were placed in the horizontal position. Thereafter thelower surface became increasingly positively charged with respectto the upper surface for at least a further 12 min. In decapitatedshoots there was no tendency whatsoever for the lower surfaceof the horizontal shoot to become positively charged with respectto the upper surface, even after 25 min in the horizontal position.The static-drop electrode system has an inherent sensitivityto reorientation in a gravitational field; a potential differencedevelops immediately after reorientation, and increases to amaximum value within the first 10 min of reorientation, regardlessof whether or not the electrodes are in contact with plant tissue.The continued increase in the potential difference measuredacross the shoots with the static-drop electrode system, andthe entire development of the potential difference measuredwith the flowing-solution electrode system, are both dependentupon the shoot apex being intact. These facts have enabled usto show that the electro-potential difference measured acrosshorizontally placed non-decapitated tissues with the static-dropelectrode system is the resultant of two distinct processes:(a) an immediate, purely physical electrical effect generatedin the electrodes themselves, and (b) a delayed geoelectriceffect which arises solely in the living tissues of the shootand which is dependent upon the apex of the shoot being intact.     

Photosynthesis of Grass Species Differing in Carbon Dioxide Fixation Pathways : VII. CHROMOSOME NUMBERS, METAPHASE I CHROMOSOME BEHAVIOR, AND MODE OF REPRODUCTION OF PHOTOSYNTHETICALLY DISTINCT PANICUM SPECIES

Panicum species of the Laxa group were investigated in a series of published reports and were found to possess C₄, C₃, and intermediate photosynthetic characteristics. Taxonomic and other relationships among these plants, however, are not clear. It was the objective of this investigation to document chromosome number, metaphase I chromosome behavior, and mode of reproduction, including abnormalities in the embryo sac, for these species.     

Enzyme Activities Associated with Carbon Dioxide Exchange in Illuminated Leaves of Hordeum vulgare L.: IV. The Effect of Light Intensity on the Carbon Dioxide Compensation Point

The carbon dioxide compensation point () was found to vary whenmeasured at increasing light intensities, in plants grown ata constant illumination. This response varied with the physiologicalage of the leaf. The also varied when measured at a constantillumination, with plants grown at different light intensities.The activity of the enzymes RuDP carboxylase, nitrate reductase,glycollate oxidase, and catalase was found to be influencedby the light intensity at which the plants were grown. A goodcorrelation was obtained between the measured and the ratioof nitrate reductase: RuDP carboxylase activities, suggestingthat nitrate reductase may be used as an indirect measure ofphotorespiration in plants receiving nitrate as the sole nitrogensource.     

Metal Interrelationships in Plant Nutrition: 2. THE RELATION OF METAL TOXICITY, MOLYBDENUM AND NITROGEN SOURCE TO CHLOROPHYLL AND MAGNESIUM CONTENT OF BEET IN SAND CULTURE

Sugar beet and spinach beet were grown in sand culture withdifferent sources of nitrogen or iron and different levels ofmolybdenum and heavy metals. Chlorophyll and some magnesiumfractions were determined. Extra molybdenum accentuated chlorosis caused by excess of Mn⁺⁺,Cr⁺⁺⁺, Zn⁺⁺, and Cu⁺⁺ and decreased it in the presence of CrO₄^––with nitrate. In the presence of these ions urea reduced chlorophyll in youngand increased it in old leaves of sugar beet. Effects of ureaand molybdenum were additive and independent. Ammonium sulphatecaused increased chlorosis of spinach beet with Mn⁺⁺ and Zn⁺⁺. The existence of an acetone-soluble magnesium fraction otherthan chlorophyll was shown. Excess Cu⁺⁺ and Zn⁺⁺ decreased totalmagnesium; Mn⁺⁺ did not. Mn⁺⁺ and Zn⁺⁺ reduced chlorophyll,increased the non-chlorophyll acetone-soluble magnesium, butreduced other fractions. Ferrous iron ipcreased total magnesiumcontent and also increased the acetone-soluble fractions inthe presence of Mn⁺⁺ and Zn⁺⁺. Extra molybdenum decreased theacetone-soluble magnesium fractions but did not affect the totalmagnesium content. Significant interactions between Mo, N, Fe, and heavy metalswere also disclosed.